The hormone-sensitive adenylate cyclase system of the murine S49 lymphoma is to be analyzed by a combination of biochemical and genetic techniques. Catecholamine-stimulated enzyme activity has been reconstituted by combination of unresponsive detergent-solubilized extracts which contain the enzyme with beta-adrenergic receptor-replete, adenylate cyclase-deficient membranes. We will attempt to resolve and purify the factors in the soluble extract that are necessary for reconstitution of the responses to hormone. There are indications that a component necessary for enzyme activity and a coupling factor are both required. The mechanisms of reinsertion of the enzyme into the membrane will be studied; this process appears to require ATP and incubation at 30 degrees. Evidence is described that indicates that at least two components are necessary to observe adenylate cyclase activity. The chemical and physical properties of these components will be studied, and initial attempts at purification will be made. Attempts will also be made to reconstitute uncoupled variants of the S49 cell by reincorporation of specific coupling factors into membranes and to create hormone responses in receptor-deficient membranes by insertion of soluble receptors for appropriate hormones.